Morphological differentiation of seven parthenogenetic Artemia (Crustacea: Branchiopoda) populations from China,with special emphasis on ploidy degrees

Parthenogenetic Artemia from seven Chinese locations with different elevations and various ploidies are characterized by phenotypic and morphometric analyses. Our findings show that the studied populations exhibit dissimilar patterns of ovisac. Four phenotypic patterns of furca are qualified and one of them is shared among di‐, tetra‐ and pentaploid Artemia. Results of discriminant analysis based on morphometric data reveal that tetra‐ and pentaploid populations are grouped together, but the Aqqikkol Lake population is clearly differentiated. Previous hypothesis/conclusion that polyploid Artemia are larger than diploids is only partly supported by the present results, which show that pentaploid and tetraploid populations are larger than the mostly diploid populations in terms of the total length, but the body size of the Aibi Lake triploids has not significant difference with the sympatric diploids and the mostly diploid Aqqikkol population that inhabit in very high altitude has the largest body size among all parthenogenetic populations. The founding confirms that body size of Artemia is following with Bergmann's rule. Microsc. Res. Tech. 79:258–266, 2016. © 2016 Wiley Periodicals, Inc.     

Comparative observations on the cyst shells of seven Artemia strains from China

The quiescent Artemia cysts of seven geographical origins in China were examined with scanning and transmission electron microscopes. SEM observations on cysts of these Artemia strains showed that the surface topography of cyst shells could be categorized into 6 types: complete smooth surface; smooth surface with sparsely distributed glabrate humps; surface with densely arranged wart-like humps that are composed of packed minute tubercles; rugged surface, with densely arranged tubercles not piling up to form larger humps; shallow-pocked surface; and surface with numerous and densely spaced pore-like fossulae. Some of the patterns were strain specific [e.g., cysts from Ga Hai (GH) are characterized by having a surface with wart-like ornaments that are composed of packed minute tubercles, rugged surface is only found in Chengkou (CK) cysts], and apparent intrastrain variation of cyst surface topography was found in Xizang (XZ), Jingyu Hu, and Xie Chi (SIN) strains. TEM studies on the ultrastructure of cyst shells revealed an apparent divergence in the structure of outer cuticular membrane (OCM) among Artemia strains. In CK, Aqqikkol Hu (AQK), SIN, and GH strains, it is a normal, asymmetrical, and multi-layered structure similar to those described in previous works. In XZ, JYH, and Lagkor Co (LGC) strains, however, the OCM is not obviously multi-layered and the borderlines between OCM and adjacent layers seem indistinct. The present results suggest that the diversity of the surface topography of Artemia cysts may be an available tool for identifying certain Artemia strains as well as for tracking the origins of some Artemia cysts, and the hypoplastic OCM may be a characteristic of the species A. tibetiana.     

Ultrastructure of cyst shell and underlying membranes of three strains of the brine shrimp Artemia (Branchiopoda: Anostraca) from South India

The cyst of Artemia has shell and membranous coverings over the embryo. The membranous coverings have special adaptive features to allow the physical changes accompanying repeated hydration and dehydration cycles that might occur and adversely influence postembryonic development. Whole and slices of cryptobiotic cysts were processed for electron microscopy to study the internal details and to compare the morphological architecture of three Artemia strains of South India. Surface topography of scanning electron microscopic (SEM) studies revealed distinct button shaped structures on the cyst of Puthalam strain. Transmission electron microscopic (TEM) studies of the cysts displayed the conventional pattern of anostracan crustaceans with outer cortex and alveolar layer, cuticular membranes, and the cytoplasmic inclusions namely nucleus, yolk droplets, lipoid bodies, and mitochondria. The prominent wavy outer cortex layer of Puthalam cysts corroborates the results of SEM studies.     

Scanning reflection electron microscopy of surface topography by diffusely scattered electrons in the scanning electron microscope

In this paper, the technique of scanning reflection electron microscopy (SREM) by diffusely scattered electrons in the scanning electron microscope is described in detail. A qualitative account of the formation of image contrast in SREM is also described. It is assumed that, for grazing geometry, forward-scattered electrons reflect from regions close to the surface, following a few scattering events within the first few atomic layers, and lose very little energy in the process. The penetration depth of the primary electrons is very limited, resulting in strongly peaked envelopes of forward-scattered electrons. It is also assumed that a surface containing topographic features presents a range of tilt angles, resulting in different reflection coefficients. Tilt contrast results because each facet has a different scattering yield, which is dependent upon local surface inclination. Full details of the instrumentation designed for SREM are described, and to illustrate the technique, results recorded from an epitaxial GaAs on GaAs crystal, Pb₂(Zr,Ta)O₆ thin film on silicon, and SiO₂ amorphous film on silicon are presented.     

Morphological characterization of Hypnaceae (Bryopsida,Hypnales): Investigating four genera from Western Himalayas by using LM and SEM techniques

The Hypnaceae is one of the largest and diversified family among the pleurocarpous mosses which consists of nearly 60 genera and 1,000 species around the world. In Pakistan, it is represented by 15 species and 8 genera. The current research article provides the detail morphological investigation of four different species (genera) of the family Hypnaceae that is, Gollania clarescens, Hypnum revolutum, Homomallium andoi, and Taxiphyllum taxirameum in the Western Himalayas, Pakistan. The research article reports the first SEM study of the leaf surfaces of the studied taxa, along with comprehensive morphological characters of the four species. The main objective of the research project is to present the comparative light and scanning electron microscopic study to discuss the morphology in detail because previously the family is just reported in different bryophyte checklists of the Western Himalayas. Based on results, morphological characters, micromorphological observations, qualitative and quantitative attributes, taxonomic keys for the studied taxa, distribution, and habitat are described. Stereoscope was used to analyze different qualitative characters, and light microscope was used to observe and measure the laminal and alar cells of the leaf. The SEM study reveals many important surface features like cell shape and cell wall. The laminal cells were mostly linear and elongated with thin cell walls. The specimens used for the SEM were air dried, so the laminal cells were somewhat constricted and concaved. The current study project will help to make the contribution in the taxonomy and morphology of this family.     

SEM analysis of enamel surface treated by Er:YAG laser: influence of irradiation distance

BACKGROUND: Depending on the distance of laser tip to dental surface a specific morphological pattern should be expected. However, there have been limited reports that correlate the Er:YAG irradiation distance with dental morphology. PURPOSE: To assess the influence of Er:YAG laser irradiation distance on enamel morphology, by means of scanning electron microscopy (SEM). METHODS: Sixty human third molars were employed to obtain discs (approximately =1 mm thick) that were randomly assigned to six groups (n=10). Five groups received Er:YAG laser irradiation (80 mJ/2 Hz) for 20 s, according to the irradiation distance: 11, 12, 14, 16, or 17 mm and the control group was treated with 37% phosphoric acid for 15 s. The laser-irradiated discs were bisected. One hemi-disc was separated for superficial analysis without subsequent acid etching, and the other one, received the phosphoric acid for 15 s. Samples were prepared for SEM. RESULTS: Laser irradiation at 11 and 12 mm provided an evident ablation of enamel, with evident fissures and some fused areas. At 14, 16 and 17 mm the superficial topography was flatter than in the other distances. The subsequent acid etching on the lased-surface partially removed the disorganized tissue. CONCLUSIONS: Er:YAG laser in defocused mode promoted slight morphological alterations and seems more suitable for enamel conditioning than focused irradiation. The application of phosphoric acid on lased-enamel surface, regardless of the irradiation distance, decreased the superficial irregularities.     

Comparison of restorative materials and surface alterations after prebiotic and probiotic beverages: A nanoindentation and SEM study

The purpose was to investigate the surface characteristics of various resin-based materials by immersing in probiotic beverages. A total of 420 disc-shaped samples (5 mm × 2 mm) were prepared from resin-based composites. Samples were divided into four groups and immersed for 10 min/day for 1 month in either a probiotic sachet, kefir, kombucha, or artificial saliva (control). Surface roughness was measured at baseline and 1 month. One sample of each of the tested materials was examined under nanoindentation to evaluate the reduced elasticity modulus and nanohardness scores. Scanning electron microscopy (SEM) was used to compare surface differences. Data were analyzed statistically using one-way ANOVA test and the significance was set at p < .05. The lowest roughness scores were observed in Z250, Estelite Bulk Fill, and HRi ENA in most of the test groups. Among conventional composites, Z250 group had the highest nanohardness and elasticity modulus scores. Among bulk-fill composites, Estelite Bulk Fill Flow had the lowest surface roughness after immersion in probiotic beverages and the highest nanohardness values. Reveal HD, as a bulk-fill group showed higher surface roughness and considerably lower nanohardness and elasticity modulus scores. Maximum height levels of samples were recorded. SEM images revealed voids and microcracks on the surfaces of test materials. Dentists may prefer Z250 as microhybrid and Estelite Bulk Fill Flow as bulk-fill composites for the restorations of patients who consume gut-friendly drinks regularly. When there are various types of materials, nanoindentation is a useful method for evaluating surface alterations and sensible comparisons.     

A SEM study of the antenna and mouthparts of Omosita colon (Linnaeus) (Coleoptera: Nitidulidae)

There are direct relationships between the behavioral mechanisms and sensilla. To obtain a better understanding of the behavioral mechanisms in Omosita colon (Linnaeus) (Coleoptera: Nitidulidae), we investigated the types, quantities, and distribution of sensilla on the antenna and mouthparts of O. colon by scanning electron microscopy. The clavate antenna comprised the scape, pedicel, and nine segment flagellomeres and had six types of sensilla, including two subtypes of sensilla chaetica (SC), three subtypes of sensilla basiconica (SB) and sensilla trichodea (ST), and one type of sensilla cavity, sensilla styloconica, and Böhm bristles (BB). The chewing mouthparts of O. colon consist of the labrum, mandible, maxillae, labium, and hypopharynx and had seven types sensilla, including two subtypes of SC and sensilla placodea, seven subtypes of SB, and one type of BB, ST, sensilla coeloconica, and sensilla campaniformia. In this research, we also deduced the relationships between the sensilla on the antenna and mouthparts and their functions.     

Thermal effects of the electron beam and implications of surface damage in the analysis of bone tissue

Electron beam interactions with specimens in the scanning electron microscope (SEM) can lead to increased surface temperatures and damage. These changes may have significant consequences in the analysis of bone tissue. An investigation was performed to measure the surface temperature changes associated with the electron beam on a thermocouple with systematic variations in operating conditions. Probe currents, magnifications, and accelerating voltages were incrementally adjusted to measure the temperature changes and to make assessments for determining optimal operating conditions for the SEM in future analyses of bone tissue. Results from this study suggest that thermal effects were minimal at lower accelerating voltages (< 20 kV), lower probe currents (< 10 nA), and lower magnifications, but surface damage may still occur during the analysis of bone tissue.     

Reproducibility and accuracy of spatial measurements from digitally captured images in the environmental scanning electron microscope

Accurate spatial measurements in a scanning electron microscope (SEM) require calibration of the magnification as a function of working distance and microscope operating conditions. This work presents the results of the calibration of an environmental SEM for the accurate spatial measurement of dimensions and areas in experiments, both for the measurement of strain in steel specimens under applied loads and the measurement of dimensional changes in timber with changes in relative humidity.     

A new method using Scanning Electron Microscopy (SEM) for preparation of anisopterous odonates

Anisopterous odonate male's secondary genitalia is a complex of several structures, among them the vesica spermalis is the most informative with important specific characters. The observation of those characters, mostly of membranous nature, is difficult in the Scanning Electron Microscope due to dehydration and metallization processes. In this contribution, we discuss a new and low cost procedure for the observation of these characters in the SEM, compatible with the most common agents used for preserving specimens.     

Morphology of the tongue dorsal surface of gilthead seabream (Sparus aurata)

The gilthead seabream is a food fish, one of the most frequently used in aquaculture. In the species of commercial interest, feeding in captivity is very important and this process is strictly related to the morphological characteristics of the oral cavity. The aim of this study is, using the standard procedures for light and scanning electron microscopy, to analyze the morphology of the tongue dorsal surface to show if relationships are present between the tongue morphology and the nutritional habits and choices of this farmed species. The main characteristic of the gilthead seabream oral cavity floor is the presence of an apical pouch, with, probably, a protective role mainly for the apical, free part of the tongue. Three zones, like in other teleosts, an apex, a body and a root, can be clearly distinguished. In the pouch foliate‐like papillae were observed, while the whole tongue is characterized by the presence of two types of papillae, respectively with a fungiform and cylindroid aspect, both randomly distributed throughout the whole dorsal surface of the tongue. Scattered and numerous taste buds, with the typical pear‐onion shape, together with small and numerous taste pores are also present, distributed throughout the tongue surface. Our results demonstrate that in the gilthead seabream important mechanic and sensory roles are carried out by specific anatomical structures. Our anatomical data could give, together with further biochemical and physiological data, an important support with the aim of improving the nutrition of aquaculture species. Microsc. Res. Tech. © 2012 Wiley Periodicals, Inc.     

Histology and ultrastructure of the testis and vas deferens in Pseudochorthippus parallelus parallelus (Orthoptera,Acrididae)

Pseudochorthippus parallelus parallelus (Zetterstedt, 1821) (Orthoptera, Acrididae) is a widespread species in Europe, and also it is localized in some regions in Turkey such as Bursa, Eski?ehir, Ankara, Bolu, Düzce, and Çank?r?. The features of the reproductive organs such as the numbers and shapes of testes and follicles can be used as taxonomical characters. For this purpose, the ultrastructural and histological features of testis and vas deferens in P. parallelus parallelus were examined with using light microscope, scanning electron microscope, and transmission electron microscope. The mature P. parallelus parallelus has two conjugated testes produce spermatozoa. Each testis is composed of numerous testis follicles in which different stages of spermatogenesis and spermiogenesis develop. First, spermatocytes are formed by the mitosis division of the germ cells at the distal end of the follicles. Then, spermatocytes form spermatids by meiosis division in the middle region of the follicles. Finally, spermatids are differentiated to spermatozoa at the proximal region of the follicles. After maturation of the spermatozoa, sperm tails come together as the sperm bundles called as spermatodesm. Each follicle is connected to vas deferens via vas efferens to discharging spermatozoa. In spite of some differences, the testes and the vas deferens in P. parallelus parallelus are highly similar to the those of other species, especially Orthopteran species.     

Postembryonic development of hard jaws (trophi) in a species belonging to the Brachionus plicatilis complex (Rotifera, Monogononta): a morphometric analysis

The presence of hard jaws (trophi), with species-specific shape and size, is a taxonomic feature of Rotifera, a group of microscopic metazoans. Since trophi are used to discriminate among species, it is important to know whether these structures change in taxonomically important ways during postembryonic development. Using both SEM and optical images, we analyzed more than 100 individuals of a single clonal lineage of a monogonont rotifer, Brachionus plicatilis, in order to describe body and trophi development after hatching. Body size, expressed as lorica width and length, was isometrically related to age of the animals only during preadult stages. Trophi size, expressed as length of the different parts, was unrelated to either age or body size. Therefore, trophi elements do not grow after hatching in B. plicatilis. Despite the dimensional invariance with age, some differences in trophi size among individuals of the same clone were recorded. No difference in left-right asymmetry of the trophi was shown; the asymmetric elements of the trophi named rami consistently had the right ramus longer than the left. This constancy is in contrast to the reported trophi asymmetries in bdelloid rotifers, in which left-right asymmetries are not constant within clonal lineages. In conclusion, we suggest that also trophi size, constant within the analyzed clone, may be used as an additional taxonomic feature to help in the discrimination of taxa within the B. plicatilis complex of cryptic species.     

The development of the modified Hurst orientation transform for the characterization of surface topography of wear particles

A modified Hurst orientation transform (HOT) method for characterization of wear particle surfaces is proposed and described in this paper. The method involves the calculation of self-affine Hurst coefficients in all directions and displays the calculated coefficient values in a form of rose plot. The calculation of individual Hurst coefficients, H, is based on the rescale range (r/s) analysis (r(d)/s∼ d ^H ). The rose plot is then used to obtain three texture surface parameters, i.e.: texture aspect ratio, texture minor axis and texture direction. The effectiveness of this modified HOT and resulting surface texture parameters was evaluated. The method was first applied to computer-generated images of isotropic and anisotropic particle fractal surfaces and then to field emission scanning electron microscope images of wear particles found in synovial joints. The ability of the surface parameters to reveal surface isotropy or anisotropy, measure roughness and determine the dominant direction of surface texture was assessed. The effects of measurement conditions such as noise, gain variations and focusing on the surface parameters were also investigated. The results demonstrate that the HOT and surface texture parameters developed can successfully be used in the characterization of wear particle surface topography. This revised version was published online in August 2006 with corrections to the Cover Date.     

Macroanatomic,light and scanning electron microscopic studies of the pecten oculi in the stork (Ciconia ciconia)

This study was undertaken to investigate the pecten oculi of stork by using macroscopic, light and electron microscopic techniques. A total of 20 eyes that were obtained from 10 storks were used. The eyes were cleaned and isolated by dissection. After various procedures, four of the pecten oculi were examined by light microscope while the other four with an electron microscope. The remaining 12 eyes were assigned for macroscopic investigation. Pecten oculi of the stork were determined as accordion‐like structures that originated from n. opticus, consisting of 15–17 plica and projecting up to 2/5 of the diameter of the bulbus oculi. Light microscopic examination revealed two types of blood vessels. Afferent–efferent vessels were larger in diamater (40–45 µm), fewer in numbers, and the capillary vessels were smaller in diamater (2–5 µm) and more in numbers. There were granules including amount of melanin pigment at the apical part of the pleats. These granules were fewer and scattered randomly on the basal part of the pleats. As a result, pecten oculi in the stork, which is a migrating bird, were determined to be similar to those of other diurnal birds. Microsc. Res. Tech. 76:963–967, 2013. © 2013 Wiley Periodicals, Inc.     

Macroanatomic,light, and electron microscopic examination of pecten oculi in the seagull (Larus canus)

The present study was conducted to determine macroanatomic characteristic as well as light and electron microscopic examination (SEM) of pecten oculi and totally 20 bulbus oculi belonging to 10 seagulls (Larus canus) were used. Pecten oculi formations consisted of 18 to 21 pleats and their shape looked like a snail. Apical length of the pleats forming pecten oculi were averagely measured as 5.77 ± 0.56 mm, retina‐dependent base length was 9.01 ± 1.35 mm and height was measured as 6.4 ± 0.62 mm. In pecten oculi formations which extend up to 1/3 of the bulbus oculi, two different vascular formations were determined according to thickness of the vessel diameter. Among these, vessels with larger diameters which are less than the others in count were classified as afferent and efferent vessels, smaller vessels which are greater in size were classified as capillaries. Furthermore, the granules which were observed intensely in apical side of the pleats of pecten oculi were observed to distribute randomly along the plica.